This invention relates to the isolation of a compound namely (xe2x88x92)-wikstromol (4,41,8-trihydroxy-3,31-dimethoxyliganan-9,91-olide) from a plant source Cedrus deodara in significant yields. This invention also identifies the use of the compound as a hepatoprotective agent.
The compound (xe2x88x92)-wikstromol as such does not find much commercial value. The structural formula of (xe2x88x92)-wikstromol is shown in FIG. 1. Lignans are widely distributed in angiosperms and gymnosperms. The range of their structures and biological activities is broad. (xe2x88x92)-Wikstromol is found to be active against p-388 lymphocyte leukemia and anti-HIV activity (M. K. Khamlach, R. Dhal and E. Brown). [Premieres syntheses totales du (+)-wikstromol, de la (xe2x88x92)-Trachelogenine, De la (xe2x88x92)-Nortrachelogenine et des lignoides apparentesxe2x80x94Tetrahedran 48, 10115-10126 (1992)].
The wood of Cedrus deodara possesses diaphoretic, diuretic and carminative properties, and it is useful in treatment of fevers, piles and pulmonary and urinary disorders. The extract of the bark is astringent and useful for fevers, diarrhea and dysentery. The oleoresin of deodar and the dark-colored oil obtained from the wood are valued for their application for ulcers and skin diseases. [Ref: Wealth of India, Vol. II, P.108-10 (1950) (published by CSIR)].
A close look at literature suggests that the lignan, secoisolariciresinalldeglycoside (SDG) which has been reported to possess a multitude of activities is isolated from flaxseed [U.S. Pat. No. 5,846,944].
P. K. Agarwal and R. P. Rastogi (Phytochemistry vol. 21, No. 6, pp. 1459-1461, 1982) reported isolation of two lignans meso-secoisolariciresinol and cedrusinin from Cedrus deodara. 
The isolation of matariesional in 0.10% yield and (+)-wikstromol in 0.124% yield is reported from Wikstroemia viridiflora (Wikstromol, a new lignan from Wikstroemia viridifloraxe2x80x94Sheela Tandon and R. P. Rastogi; Phytochemistry 1976, vol. 15, pp. 1789-1791).
The extraction of the lignans and other constituents namely (xe2x88x92)-nortrachelogenin, carinol and carissanol from Carissa edulis is reported by Hans Achenbach, Reiner Waibel and Ivan Addae-Mensah, in Phytochemistry, Vol.22, No.3, pp.749-753, 1983.
There is a considerable amount of epidemiological evidence indicating an association between diets rich in fruits and vegetables and a decreased risk of cardiovascular disease and certain forms of cancer. It is generally assumed that the active principles contributing to these protective effects are nothing but antioxidant phytochemicals.
Recent research has been directed to find out the phytochemicals from plant sources and to highlight the role of polyphenolic compounds of plant materials as antioxidants, antimutagenic, anti inflammatory, antiatherosclerotic, antidiabetic, antihepatotoxic and antimicrobial agents. [An overview of flax lignans is given by Neil D. Westcott and Alister D. Muir, Crop Utilization Section, Saskatoon Research Centre, Agriculture and Agri-Food Canada, 107, Science Place, Saskatoon, SK S7N 0X2, Canada in vol. IIxe2x80x94January 2000 xe2x80x94inform]. The alcoholic extract of stem C. deodara was found to have anti-cancer activity. [See Medicinal Plants of India (ICMR) Vol. I, 1976, pp.214 and 215].
Accordingly, the applicants conducted a detailed study on principles from Cedrus deodara and this investigation led to the isolation of active principles, namely, wikstromol. These compounds although hitherto isolated from Wikstromia spp. were in low yields. Cedrus deodara, hence, is a new source for the lignans and its presence in this taxon in significantly high yields makes this invention more important.
The main object of the invention is to provide novel compositions containing (xe2x88x92)-wikstromol which are useful as a hepatoprotective agent.
Yet another object of the invention is to provide methods for the isolation of (xe2x88x92)-wikstromol from Cedrus deodara. 
Accordingly, the invention provides novel compositions containing (xe2x88x92)-wikstromol which are useful as hepatoprotective agents. The invention further provides method for the isolation of (xe2x88x92)-wikstromol from Cedrus deodara. 
Accordingly, the invention provides a composition comprising an effective amount of (xe2x88x92)-wikstromol together with or associated with an additive and useful as a hepatoprotective agent.
In an embodiment, the additive is selected in such a manner that it does not affect or interfere with the efficacy of the active principles of the composition. The additives are such that they enhance and do not retard the activity of the active ingredient, i.e. (xe2x88x92)-wikstromol.
In another embodiment, the additive is selected from nutrients such as carbohydrates, sugar, proteins and pharmaceutically acceptable carriers.
In still another embodiment, the ratio of (xe2x88x92)-wikstromol with the additive is in the range between 0.4:10 to 2.0:10.
In yet another embodiment, (xe2x88x92)-wikstromol is present in an amount of 250-300 mg.
Further, the invention provides a process for the isolation of (xe2x88x92)-wikstromol from the Cedrus deodara, said process comprising the steps of:
g) extraction of the pulverized plant parts of Cedrus deodara with solvents to remove the essential oils;
h) concentrating the extract under vacuum to obtain a residue;
i) adding ethyl acetate to the residue obtained in step (b);
j) separating the solvents by conventional methods;
k) subjecting the residue to a first elution with about 3% methanol in chloroform to obtain (xe2x88x92)-matairesinol; and
l) subjecting the residue of step (e) to a second elution with about 5% methanol to obtain (xe2x88x92)-wikstromol.
The solvents used in step (a) are hexane and chloroform.
In another embodiment, the plant parts of Cedrus deodara such as bark and leaves are used for extraction.
In yet another embodiment, the wasted plant parts of Cedrus deodara are employed for isolation of the said compound. Preferably, the waste left, after extraction of essential oil from the plant parts, is used in the process.
Accordingly, the compositions prepared can be used as hepatoprotective treatment agents, wherein an effective amount of (xe2x88x92)-wikstromol is administered to a subject in need thereof.
(xe2x88x92)-Wikstromol may be administered together with or in combination with therapeutically acceptable additives. The effective amount of (xe2x88x92)-wikstromol that may be administered to a subject can be readily determined by a person skilled in the art. However, it is recommended that the dosage of (xe2x88x92)-wikstromol administered may be in the range of 250 to 300 mg per dose, twice a day. While it is possible to administer the composition in many routes, the oral route achieves the desired, best results.
Compositions employing (xe2x88x92)-wikstromol may be prepared by conventional methods as may be known in the art. The compositions may be in the form of tablets, capsules or syrups, etc. Suitable additives as may be known in the art may be selected for the preparation of these compositions.
In essence, the focus of the invention is to provide methods for using (xe2x88x92)-wikstromol for the preparation of compositions useful as hepatoprotective agents.
The heartwood of Cedrus deodara finds extensive use in the essential oil industry. The oil named xe2x80x98cedar woodxe2x80x99 oil finds applications in flavor and fragrances. The heartwood powder, after extraction of the essential oil, is a by-product and waste.
This invention relates to isolation and purification of the compound (xe2x88x92)-wikstromol, which is useful as a hepatoprotective agent.
The present invention relates to the isolation of a compound, namely (xe2x88x92)-wikstromol (4,41,8-trihydroxy-3,31-dimethoxyliganan-9,91-olide) from a new plant, Cedrus deodara. This invention also relates to a new use of the compound, i.e., as a hepatoprotective agent.
The present invention embodies the isolation of (xe2x88x92)-wikstromol, an antioxidant principle from an entirely new source and its superior antihepatotoxic property compared with known biologically proved anti hepatotoxic agents.